JP2024032817A - Brown fat/beige fat cell activator containing D-allulose as an active ingredient - Google Patents

Abstract

[Object] To provide a brown adipocyte or beige adipocyte activator and a food for brown adipocyte or beige adipocyte activation, which contain D-allulose as an active ingredient. [Solution] The weight and area morphology of brown adipocytes or beige adipocytes containing D-allulose as an active ingredient, whose function is activated by D-allulose, for example, the expression level of an activation indicator is increased. target increaser. Brown adipocytes or beige adipocytes in which the expression level of the activation indicator is increased are brown adipocytes or beige adipocytes in which UCP-1 expression is promoted or enhanced and the expression level is increased. An energy consumption promoter containing D-allulose as an active ingredient, which is an agent for morphologically increasing the weight and area of brown fat cells or beige fat cells whose expression levels of activation indicators are increased by D-allulose. [Selection diagram] Figure 16

Description

The present invention relates to a brown fat/beige fat cell activator. More specifically, the present invention relates to a brown fat/beige fat cell activator containing D-allulose.

Mammalian adipose tissue is broadly classified into two types, white adipose tissue and brown adipose tissue, depending on its function and histological characteristics. White fat cells present in subcutaneous and visceral white adipose tissue store excess energy in the body as fat. On the other hand, brown fat cells are responsible for burning fat and producing heat. The physiological role of brown adipose tissue is completely opposite to that of white adipose tissue; it is a site that consumes and dissipates energy as heat when stimulated by sympathetic nerves. Brown adipose tissue contains brown adipocytes, which are characterized by having multilocular lipid droplets and abundant mitochondria. protein 1)] has the function of dissipating energy as heat. Brown adipose tissue is particularly abundant in newborns and hibernating animals. Its main function is to help animals and newborns generate body heat without shivering. In contrast to white adipocytes, which contain a single lipid droplet, brown adipocytes contain iron, which gives them a brown color, and contain numerous small droplets and a much higher number of mitochondria. It is. Brown adipose tissue requires more oxygen than most tissues, so it has more capillaries than white adipose tissue. Previously, it was thought that brown fat cells are abundant in the body during childhood, but are lost as adults. However, recent research has begun to use PET scans, and it has become clear that brown fat cells remain and function in areas such as the neck and shoulders even in adults.

It is known that catechins contained in tea, etc., and capsaicin contained in chili peppers, etc. have the effect of activating brown adipose tissue, and there is a desire for further substances that are highly effective in activating brown adipose tissue. However, there have been no reports that rare sugars have an activating effect on brown fat-like cells.

International Publication No. 2010/113785

Applied Glycoscience Vol. 5 No. 1 44-49 (2015) Iichiro Shimomura et al., Journal of the Japanese Society of Internal Medicine, 93(4), 655-661, 2004 Ikeda K. et al., TEM, 2018;29:191-200

In addition to burning energy to warm the body at low temperatures, brown fat cells are thought to increase energy consumption and make it easier to burn fat.
In other words, if we can find a way to activate brown fat cells, it could become a new treatment for controlling obesity and type 2 diabetes.
An object of the present invention is to provide a preparation that has a high activation effect on brown fat cells.

Furthermore, an object of the present invention is to provide a composition that exhibits an effect of promoting UCP-1 expression and/or an effect of promoting brown adipocyte differentiation, which can be ingested over a long period of time and continuously without the problem of side effects. do.
Furthermore, the present invention aims to provide a brown adipocyte activator and a food for brown adipocyte activation, which contain D-allulose as an active ingredient.

In addition to burning energy to warm the body at low temperatures, brown fat cells are thought to increase energy consumption and make fat easier to burn. Focusing on this function of cells, in other words, if we can find a way to activate brown fat cells, it may become a new treatment to control obesity and type 2 diabetes.As a result of intensive research, we have found that brown fat cells can be browned by administration of rare sugars. We have obtained knowledge regarding fat cell activation and associated weight loss, leading to the invention of a brown fat cell activator containing D-allulose as an active ingredient.
In the present invention, "activation of brown fat cells" refers to an action that promotes induction of differentiation of brown fat-like cells (beige fat cells) in white fat tissue, and/or an action that promotes induction of differentiation of brown fat cells (beige fat cells) into white fat tissue, and/or an action that promotes differentiation of brown fat cells (beige fat cells) into white fat tissue. It refers to the action of promoting energy consumption in the body, and can also be called the activation of brown fat/beige fat cells.

In addition to burning energy to warm the body at low temperatures, brown fat cells are thought to increase energy consumption and make fat easier to burn. Focusing on this function of cells, in other words, if we can find a way to activate brown fat cells, it may become a new treatment to control obesity and type 2 diabetes.As a result of intensive research, we have found that brown fat cells can be browned by administration of rare sugars. We have obtained knowledge regarding fat cell activation and associated weight loss, leading to the invention of a brown fat cell activator containing D-allulose as an active ingredient.
In the present invention, "activation of brown fat cells" refers to an action that promotes induction of differentiation of brown fat-like cells (beige fat cells) in white fat tissue, and/or an action that promotes induction of differentiation of brown fat cells (beige fat cells) into white fat tissue, and/or an action that promotes differentiation of brown fat cells (beige fat cells) into white fat tissue. It refers to the action of promoting energy consumption in the body, and can also be called the activation of brown fat/beige fat cells.
The gist of the present invention is the following agents (1) to (4) for morphologically increasing the weight and area of brown adipocytes or beige adipocytes.
(1) An agent for morphologically increasing the weight and area of brown fat cells or beige fat cells whose functions are activated by D-allulose, which contains D-allulose as an active ingredient.
(2) The brown adipocytes or beige adipocytes according to (1) above, wherein the functionally activated brown adipocytes or beige adipocytes are brown adipocytes or beige adipocytes in which the expression level of an activation indicator is increased. Agent for morphologically increasing the weight and area of beige adipocytes.
(3) Brown adipocytes or beige adipocytes in which the expression level of the activation indicator has increased are brown adipocytes and beige adipocytes in which UCP-1 expression has been promoted and the expression level has increased; ) The agent for morphologically increasing the weight and area of brown adipocytes or beige adipocytes.
(4) Claim 2, wherein the brown adipocytes or beige adipocytes in which the expression level of the activation indicator is increased are brown adipocytes or beige adipocytes in which UCP-1 expression is enhanced and the expression level is increased. The agent for morphologically increasing the weight and area of brown adipocytes or beige adipocytes according to .

Moreover, the gist of the present invention is the following energy consumption promoter (5) or (6).
(5) An energy consumption promoter containing D-allulose as an active ingredient, which is an agent for morphologically increasing the weight and area of brown fat cells or beige fat cells whose expression levels of activation indicators are increased by D-allulose.
(6) Brown adipocytes or beige adipocytes in which the expression level of the activation indicator has increased are those in which energy consumption is promoted due to the expression of genes involved in thermogenesis, and the expression level has increased in brown adipocytes or beige adipocytes. An energy consumption promoter according to (5) above.

Moreover, the gist of the present invention is the following UCP-1 expression enhancer (7).
(7) UCP-1 expression enhancement consisting of an agent that morphologically increases the weight and area of brown adipocytes or beige adipocytes, which contains D-allulose as an active ingredient and whose expression level of activation indicators is increased by D-allulose. agent.

Further, the gist of the present invention is the composition for morphologically increasing the weight and area of brown adipocytes and beige adipocytes according to (8) to (12) below.
(8) A composition for morphologically increasing the weight and area of brown adipocytes or beige adipocytes, comprising the agent according to any one of (1) to (7) above.
(9) A composition for morphologically increasing the weight and area of brown adipocytes or beige adipocytes in which the expression level of an activation index is increased by D-allulose, which contains D-allulose as an active ingredient.
(10) Brown adipocytes or beige adipocytes in which the expression level of the activation indicator has increased, energy consumption is promoted due to the expression of genes involved in thermogenesis, and brown adipocytes and beige adipocytes in which the expression level has increased; A composition for morphologically increasing the weight and area of brown adipocytes or beige adipocytes according to (9) above.
(11) The above-mentioned ( 9) The composition for morphologically increasing the weight and area of brown adipocytes or beige adipocytes.
(12) The weight and area of brown fat cells or beige fat cells according to any one of (9) to (11) above, which is a food composition for morphologically increasing the weight and area of brown fat cells or beige fat cells. A composition for the morphological increase of.

According to the present invention, a brown fat/beige fat cell activator, an energy consumption promoter, a UCP-1 expression enhancer, etc. containing D-allulose as an active ingredient has a high activation effect on brown fat/beige fat cells. A formulation can be provided.

Furthermore, the present invention provides a composition and a food composition containing a brown fat/beige fat cell activator, an energy consumption promoter, and a UCP-1 expression enhancer containing D-allulose as an active ingredient. can. In addition, compositions for activating brown fat/beige fat cells containing D-allulose as an active ingredient, compositions for activating brown fat/beige fat cells by enhancing expression of UCP-1, food compositions, etc. To provide a composition or a food composition that exhibits an effect of promoting UCP-1 expression and/or an effect of promoting brown fat/beige fat cell differentiation, which has no side effects and can be ingested over a long period of time. Can be done.

FIG. 2 is a diagram showing changes in body weight and blood sugar levels in an experiment of Example 1 using 6-week-old mice. FIG. Regarding the effect of D-allulose on mouse body weight, high-fat diet (HFD) increased body weight over 8 weeks of observation. When the HFD group was made to drink water containing 2% D-allulose, a decrease in body weight was observed. In addition, blood sugar levels were measured over time during 8 weeks. In the HFD group, blood sugar levels were higher than in the normal group. Blood glucose levels in the HFD+D-allulose group were lower than in the HFD group. 1 is a drawing showing the size and weight of subscapular brown fat cells and body weight in the experiment of Example 1 using 6-week-old mice. Furthermore, it is a drawing (an image of an electron micrograph) showing the results of pathological examination of brown adipose tissue in Group 3. Compared to the normal diet group, in the HFD group, fat deposits were observed in the BA, which turned into white fat. On the other hand, in the HFD group + 2% D-allulose group, the brown adipose tissue was morphologically comparable to that in the normal diet group, and it was found that the brown adipose tissue that had turned into white fat had recovered to normal brown adipose tissue. . FIG. 2 is a drawing showing the expression level of UCP-1 and the activation of brown adipocytes in the experiment of Example 1 using 6-week-old mice. FIG. In this example, UCP-1 was increased in the HFD group. It was predicted that ingesting a large amount of fat would induce UCP-1 as a body defense response, leading to calorie consumption. Administration of D-allulose further induced UCP-1. When considered in conjunction with pathological tissue, the induction of UCP-1 in the HFD group is a response to a pathological situation, and in the HFD + 2% D-allulose group, brown adipose tissue is induced due to the normal morphology. It is presumed that this is the activation of UCP-1 as a result of its activation. FIG. 2 is a drawing showing activation of brown adipocytes and the expression level of PPAR-α in the experiment of Example 1 using 6-week-old mice. FIG. PPAR-α is an indicator of brown adipocyte activation. In this example, PPAR-α was induced in the HFD group compared to the normal group, but this is thought to be a biological reaction due to excessive lipid intake. In the HFD group + 2% D-allulose group, PPAR-α was more strongly induced by the intake of D-allulose, suggesting that brown fat cells were activated. 1 is a diagram showing the expression level of PGC-1α and the activation of brown adipocytes in the experiment of Example 1 using 6-week-old mice. In this example, PGC-1α was induced in the HFD group compared to the normal group, but this is thought to be a biological reaction due to excessive lipid intake. In the HFD + 2% D-allulose group, PGC-1α was more strongly induced by D-allulose intake, and the induction of PGC-1α, which is a coactivator of PPAR-α, is due to UCP-1, which is located in the downstream region. This is thought to be one of the reasons why brown fat cells are activated. 2 is a diagram showing the change in body weight in the human clinical trial of Example 2. 1 is a diagram illustrating various adipose tissue-derived physiologically active substances (adipocytokines) and their effects (excerpted from Non-Patent Document 2). It is known that adipocytes are cells that not only serve as a storage source of energy but also produce and release numerous cytokines (physiologically active substances). Cytokines produced and released from fat cells are called adipocytokines. Adipocytokines are also deeply involved in metabolic syndrome. TNF (tumor necrosis factor)-α causes phosphorylation of IRS-1, resulting in insulin resistance. Adiponectin is known to have suppressive effects on obesity, diabetes, and arteriosclerosis, and is interpreted as a good substance. It is known that this value decreases in obesity and diabetes. FIG. 2 is a diagram illustrating the action of adiponectin and the possibility of therapeutic application. As is well known, TNF-α is secreted from adipocytes and is famous as a cytokine that induces insulin resistance. It is also known to suppress adiponectin, a good adipocytokine. It is presumed that the administration of D-allulose (D-psicose) in this example lowers TNF-α, which improves insulin resistance and improves blood sugar control.

3 is a diagram illustrating the experimental procedure (protocol) of Example 3. FIG. Each group consists of 5 mice and is divided into 2 groups. Both groups of 8 week old mice are fed a high fat diet (HFD). Four weeks after HFD loading, group 1 continued on HFD. In another group, in addition to HFD, D-allulose (0.2 mg/g body weight/day) was administered into the stomach using a probe. Body weight, food intake, water consumption, and blood sugar levels will be monitored until the 10th week. FIG. 2 is a diagram showing the change in body weight of mice in Example 3 when fed a normal diet and a high-fat diet (HFD). 2 is a diagram showing the change in body weight (BW) when D-allulose was administered to mice fed a high-fat diet (HFD) in Example 3. In the group to which D-allulose was added after the 4th week, body weight decreased significantly. 12 is a diagram showing changes in fasting blood sugar levels in Example 3. In the D-allulose addition group, a tendency towards a decrease in fasting blood sugar levels was observed. 3 is a diagram showing the transition of blood sugar levels in a glucose tolerance test after 10 weeks in Example 3. FIG. In the glucose tolerance test, the fasting blood sugar level and the 90-minute and 120-minute post-load blood sugar levels were significantly lower. FIG. 7 is a drawing showing the amount of food intake (right side diagram) and water consumption (left side diagram) during the research period in Example 3. There were no significant changes in food intake or water consumption between the two groups, and there were no obvious differences. FIG. 2 is a diagram showing the weight and area of brown adipose tissue (BAT) standardized by body weight when D-allulose was administered to mice loaded with a high-fat diet (HFD) in Example 3. FIG. In the group supplemented with D-allulose, the weight and area of brown adipose tissue (BAT) increased. This is a drawing explaining the distribution and function of brown adipose tissue (Burn energy for thermogenesis) (excerpted from Non-Patent Document 3). It is a drawing showing the expression level of UCP-1 mRNA in beige adipocytes and brown adipocytes in mice fed with a high-fat diet (HFD) and administered with D-allulose. The drawing on the left shows that the expression of UCP-1 is significantly increased in beige adipocytes by administration of D-allulose. The drawing on the right shows that UCP-1 expression is increased even in classical brown adipose tissue. It is a drawing showing the expression level of UCP-1 protein in beige adipocytes and brown adipocytes in mice fed with a high-fat diet (HFD) and administered with D-allulose. In beige adipocytes, administration of D-allulose significantly increases the expression of UCP-1 protein. UCP-1 protein expression is also increased in classical brown adipose tissue. Regarding induction of beige adipocytes and activation of brown adipocytes by D-allulose administration, genes involved in heat production, UCP-1 and Prdm16, gene Pgcl-α, which reflects the function of mitochondria, an organelle involved in heat production, It is a drawing showing the expression level of Tfam and PPARγ, a gene related to adipose differentiation. For experiments using cells (establishing a cell line derived from a single clone from BAT in the adult supraclavicular fossa and using a model for the study of differentiation induction into beige adipocytes), the BAT-protocol and WAT-protocol are used. 2 are drawings explaining a protocol for inducing beige adipocytes. In an experiment using cells, the BAT-protocol and the WAT-protocol are diagrams explaining protocols in which D-allulose is added to the protocols for inducing beige adipocytes. FIG. 2 is a diagram (an image of an electron micrograph) showing by fat staining that addition of D-allulose to the differentiation induction protocol promotes induction into beige adipocytes. 1 is a drawing showing the results of examining the expression of various markers (UCP-1, Prdm16, Pgc-1α, tfam, cox8b, PPARγ) as an evaluation of beige adipocytes induced by a differentiation induction protocol. The WAT-protocol has a stronger effect on the induction of browning marker genes (UCP-1, Pgc-1α, cox8b). Using the WAT-protocol, D-allulose enhances the expression of Prdm16, Pgc-1α, and PPARγ. In both differentiation protocols, D-allulose enhanced PPARγ expression, suggesting that D-allulose promotes adipogenesis. As an evaluation of beige adipocytes induced by the protocol, the expression of UCP-1 and PPARγ was examined in the presence and absence of D-allulose, and this drawing shows the results. It was shown that the presence of D-allulose increased UCP-1 and PPARγ, promoting the induction of beige adipocytes. Compared to the BAT-protocol, the WAT-protocol has a stronger effect on the induction of UCP-1 protein expression. Using the WAT-protocol, D-allulose slightly increases UCP-1 protein expression.

[Brown fat cells]
In the present invention, brown fat cells refer to both classical brown fat cells formed during the fetal period and brown fat-like cells (sometimes called beige fat cells or bright cells) that are induced to differentiate into white adipose tissue. Point. That is, in the present invention, not only classical brown adipocytes in the narrow sense but also brown-like adipocytes induced in white adipose tissue are referred to as brown adipocytes. Furthermore, brown fat cells induced into white adipose tissue are sometimes referred to as beige fat cells or brown fat cells.

[White fat cells]
White adipocytes have large unilocular lipid droplets and have little cytoplasm. On the other hand, brown fat cells have small multilocular lipid droplets, and a large number of mitochondria exist around these multilocular lipid droplets, giving them a distinctive brown color and rich in sympathetic nerves and blood vessels. It has the following morphological and histological characteristics. Therefore, white adipocytes and brown adipocytes can be distinguished by morphologically and histologically observing the cells. Another difference is that white fat stores energy, whereas brown fat cells consume and dissipate energy as heat. Furthermore, brown adipocytes have higher energy metabolism than white adipocytes, and increase glucose uptake to release energy as heat. Therefore, the presence of brown fat cells can be evaluated, for example, by measuring the accumulation of 18F-labeled glucose using PET (positron emission tomography). Furthermore, in brown adipocytes, a 33 kDa protein called uncoupling protein 1 (UCP-1) is specifically expressed in the inner mitochondrial membrane of the cell, so we measured the expression of UCP-1 mRNA and UCP-1 protein. By doing so, the presence of brown fat cells can be confirmed.

[Beige fat cells]
Morphologically, beige adipocytes are similar to brown adipocytes, have intracellular multilocular lipid droplets, and are rich in mitochondria that express the specific protein UCP-1. This is in contrast to white adipocytes, which have unilocular lipid droplets and lack cytoplasm. Looking at their functional characteristics, unlike white adipocytes, which store surplus energy as neutral fat, brown adipocytes and beige adipocytes generate heat by uncoupling oxidative phosphorylation by UCP-1. . In these respects, beige adipocytes can be said to be similar to brown adipocytes, but in the following respects they are more similar to white adipocytes. First, regarding the location of their existence, in mice, brown adipocytes are localized by forming cell clusters between the shoulder blades and around the kidneys, whereas beige adipocytes are induced and scattered in white adipose tissue such as in the inguinal region. Appear. This phenomenon is called browning of white fat. Next, regarding the developmental origin, brown adipocytes are derived from muscle progenitor cells that express Мyоgenic factor 5 (Мyf5), which is common to skeletal muscle, whereas beige adipocytes, like white adipocytes, are Мyf5 negative and are plate-derived. It is derived from preadipocytes that express growth fact receptor α (PDGFRα) and smooth muscle actin (SMA). In this way, beige adipocytes have both similar and contradictory characteristics to brown adipocytes and white adipocytes, so they are not simply white adipocytes with changed properties, but are considered to be a third type of adipocyte. It will be done.
Brown adipocytes and beige adipocytes are specialized adipocytes that generate heat in response to cold exposure, contributing to maintaining body temperature in cold environments. The thermogenic and energy consuming activities of these fat cells are expected to be useful not only for regulating body temperature but also for preventing obesity and metabolic diseases. Brown adipocytes and beige adipocytes express uncoupling protein (UCP-1) and have thermogenic ability in common, but it has been found that their cell origins and functional control mechanisms are different. ing. In particular, the fact that brown adipose tissue (BAT) in human adults is mainly composed of beige adipocytes is considered important in searching for obesity prevention methods targeting BAT.

In the present invention, activation of brown adipocytes refers to an action that promotes induction of differentiation of brown adipose-like cells (beige adipocytes) into white adipose tissue, and/or an effect that promotes the induction of differentiation of brown adipocytes (beige adipocytes) into white adipose tissue, and/or energy generation in beige adipocytes and brown adipocytes. Refers to the effect of promoting consumption. Activation of brown fat cells may increase metabolism, promote energy expenditure, and convert fatty acids into heat energy, especially in beige and brown fat cells. The conversion of fatty acids to thermal energy may also be carried out by UCP-1. Note that, as mentioned above, activation of brown fat cells is sometimes expressed as activation of brown fat/beige fat cells.

The brown fat cell activator refers to a substance that has the above-mentioned effect of activating brown fat cells. As mentioned above, the presence of brown fat cells and/or an increase in the energy metabolism of brown fat cells can be determined by measuring UCP-1 mRNA expression and UCP-1 protein, observing cells histologically, and 18F. Evaluation can be made by measuring the accumulation of labeled glucose using PET, by measuring cold-induced thermogenesis, etc. In addition, induction of differentiation of brown fat-like cells (beige fat cells) in white adipose tissue is sometimes referred to as reactivation of brown fat cells.

The brown fat cell activator of the present invention can promote energy metabolism through thermogenesis. Energy metabolism includes basal metabolism, exercise, daily activities, and heat production. Approximately 60% of the daily energy expenditure is due to basal metabolism, approximately 5% is due to exercise, approximately 24% is due to daily life activities, and approximately 10% is due to thermogenesis. Many attempts have been made to increase basal metabolic rate by increasing muscle mass, and to increase energy metabolism by exercising and being active in daily life activities. On the other hand, energy consumption due to thermogenesis allows the body to respond to rapid temperature changes, and increases, for example, due to cold stimulation. It is also known that heat production is increased by food intake, and that heat is produced to generate fever in response to infections, inflammation, etc. Therefore, in the present application, energy consumption due to thermogenesis refers to metabolic energy consumption consumed for heat production in response to cold stimulation, meal intake, etc., which is not based on basal metabolism, muscle exercise, etc. The brown fat cell activator of the present application may increase heat production in beige fat cells and brown fat cells and promote energy metabolism, particularly by stimulating sympathetic nerves. Such thermogenesis may occur by converting fatty acids into thermal energy, and the conversion from fatty acids to thermal energy may be performed by UCP-1.
The brown fat cell activator, energy consumption agent, etc. of the present invention, and compositions containing them may enhance energy metabolism not in the liver or muscle but in brown fat tissue, and may enhance UCP-1 expression. There is also.

The content of D-allulose in the brown fat cell activator or UCP-1 expression enhancer of the present invention is not particularly limited, but it is preferably incorporated in an amount of 0.3 to 50 g per day in terms of weight. and may be blended in an amount of 0.3 to 5 g, 0.5 to 3 g, 1.5 to 4.5 g, 0.5 to 5 g, 0.5 to 50 g, or 0.5 to 20 g, etc., depending on the case. There is also. Furthermore, in order to enhance the effects of the present invention, other brown fat cell activators, body temperature decrease inhibitors, energy consumption promoters, UCP-1 expression enhancers, metabolism promoters, etc. may be used in combination.

The intake amount of D-allulose is not particularly limited, but for a human weighing 60 kg, the amount is 0.3 to 50 g in terms of weight per day, and in some cases, 0.3 to 5 g, 0.5 g. It may be preferable to take amounts such as ~3g, 1.5-4.5g, 0.5-5g, 0.5-50g, or 0.5-20g.

The method for producing a brown fat cell activator of the present invention may include a step of producing D-allulose. The form of D-allulose used in the present invention is arbitrary as long as it does not impair the effects of the present invention, as described above. Currently, the common method for obtaining the rare sugar D-allulose is to treat fructose with an enzyme (epimerase). It can also be obtained by a method of treating rare sugar syrup or isomerized sugar as a raw material in a system in which one or more selected from the group consisting of a basic ion exchange resin, an alkali, and a calcium salt is present (Patent Document 1). Mainly produced to contain D-allulose and D-allose, but also contains 0.5-17% by mass of D-allulose, 0.2-10% by mass of D-allose, and other unidentified rare sugars. It will be done. The rare sugar syrup (trade name: Rare Sugar Sweet) produced in this way has glucose and fructose as its main components, and contains 5.4 g/100 g of D-psicose, 5.3 g/100 g of sorbose, 2.0 g/100 g of tagatose, and 1.0 g/100 g of allose. 4g/100g and mannose 4.3% (Non-Patent Document 1). Currently, rare sugars such as D-psicose can be produced using enzymatic methods, alkaline methods, etc., but it has also been found that they are also contained in plants such as leaves of Zinna.

The present invention also provides a composition containing the above-mentioned brown fat cell activator (brown fat/beige fat cell activator), hypothermia inhibitor, energy consumption promoter, or UCP-1 expression enhancer. . The composition of the present invention can be in various forms such as powder, liquid, solid, granule, tablet, paste, gel, emulsion, cream, sheet, spray, and foam.

The food composition of the invention may be a powder, a drink, or a tablet. The food composition of the present invention can also be used in dry powders, beverages such as tea and soft drinks, tablets and capsules such as supplements, processed foods such as retort foods, luxury foods such as desserts, seasonings, dairy products, oils and fats. It may be a processed product or the like, and may be in various forms such as powder, liquid, solid, granule, granule, paste, or gel. Furthermore, the food composition of the present invention includes not only food for humans but also feed for other animals such as livestock.

The content of the brown fat cell activator (brown fat/beige adipocyte activator) in the composition of the present invention is not particularly limited, but the dosage is D-allulose (D-allulose) for adults when administered orally. It is preferable to take 0.3 to 50 g of psicose per day, but the amount can be increased or decreased as appropriate depending on age and symptoms. The daily dose of the brown fat cell activator, energy consumption promoter, or UCP-1 expression enhancer of the present invention may be administered once a day, or divided into two or three times a day at appropriate intervals. It is preferable to administer the drug before, after, or with a meal. In the composition containing D-allulose (D-psicose) of the present invention, D-allulose (D-psicose) is contained in the composition in an amount of 0.1 to 50% by weight. Preferably it is 0.5 to 30% by weight, more preferably 1 to 10% by weight. If D-allulose (D-psicose) is less than 0.1% by weight in the composition, the brown fat cell activation effect will not be sufficient. Moreover, if D-allulose (D-psicose) exceeds 50% by weight in the composition, it is not preferable from an economical point of view.
In addition, the composition of the present invention has a brown fat cell activation effect, a body temperature drop suppression effect, an energy consumption promotion effect, etc. in order to further enhance the brown fat cell activation effect, the body temperature fall suppression effect, the energy consumption promotion effect, etc. Certain other substances may also be added.

Furthermore, in the composition of the present invention, additives can be arbitrarily selected and used in combination as necessary. Excipients and the like can be included as additives.

Any excipient may be used as long as it is commonly used in the desired form, such as starches such as wheat starch, rice starch, corn starch, potato starch, dextrin, and cyclodextrin, crystalline cellulose, Examples include saccharides such as lactose, glucose, sugar, reduced maltose, starch syrup, fructooligosaccharides, and emulsified oligosaccharides, and sugar alcohols such as sorbitol, erythritol, xylitol, lactitol, and mannitol. These excipients can be used alone or in combination of two or more.

In addition, the composition of the present invention may contain other ingredients as necessary, such as colorants, preservatives, thickeners, binders, disintegrants, dispersants, stabilizers, gelling agents, and antioxidants. , surfactants, preservatives, pH adjusters, oils, powders, colorants, water, alcohols, thickeners, chelating agents, silicones, antioxidants, ultraviolet absorbers, humectants, fragrances, various medicinal ingredients , preservatives, pH adjusters, neutralizing agents, and other known agents can be appropriately selected and used.

The brown fat cell activator (brown fat/beige fat cell activator) of the present invention can induce differentiation of brown fat-like cells (beige fat cells) in white adipose tissue, and/or beige fat cells and brown fat cells. Obesity can be prevented and/or suppressed by promoting metabolism through the approach of activating fat cells and promoting whole-body energy consumption, particularly through thermogenesis. Therefore, it is also effective in treating and/or preventing diseases caused by obesity. Additionally, by activating brown fat cells and increasing cold-induced heat production, it is possible to prevent the body from getting cold, and by increasing meal-induced heat production, it is possible to enhance energy metabolism from meals. Furthermore, regarding the safety of D-allulose (D-psicose), the active ingredient of the present invention, it is contained in various foods that we eat (cola, castella, maple syrup, etc.) For these reasons, rather than the substance being ingested, it is thought that there is no major risk in ingesting the rare sugar D-allulose (D-psicose).

Next, the present invention will be explained in more detail with reference to Examples. Note that the present invention is not limited to this.

[How to check brown fat cells]
The presence of brown fat cells can be confirmed by a known method. Examples include staining with a fluorescent dye that can detect lipid droplets in cells, and detection of gene products (mRNA or proteins) expressed in brown fat cells. Fluorescent dyes that can detect lipid droplets in cells include Oil Red O, BODIPY, and the like. Gene products expressed in brown fat cells include UCP-1, CIDEA, PGC-1α, PPAR-α, and the like. Among them, UCP-1 is a gene specifically expressed in brown adipocytes, encodes a mitochondrial inner membrane protein that uncouples oxidative phosphorylation, and is thought to play a fundamental role in the function of brown adipocytes. It is one of the particularly preferred indicators of brown fat cells.

In the Examples of the present invention, activation of brown adipocytes was performed by evaluating the expression levels of UCP-1, PPAR-α, and PGC-1α.

The terms used in the embodiments of the present invention will be briefly explained. See FIG. 8 showing various adipose tissue-derived physiologically active substances (adipocytokines) and their effects (extracted from Non-Patent Document 2) and FIG. 9 showing the effects and therapeutic applications of adiponectin.
[BAT]
Brown adipose tissue (BAT) or brown adipose tissue is one of two types of fat or adipose tissue found in mammals. The other type is white adipose tissue. Brown adipose tissue is particularly abundant in newborns and hibernating animals. Its main function is to help animals and newborns generate body heat without shivering. In contrast to white adipocytes, which contain a single lipid droplet, brown adipocytes contain iron, which gives them a brown color, and contain numerous small droplets and a much higher number of mitochondria. It is. Because brown adipose tissue requires more oxygen than most tissues, brown adipose tissue also has more capillaries than white adipose tissue.
When noradrenaline binds to β3 receptors on brown fat cells, UCP-1 (uncoupling protein) is produced, causing uncoupling in the mitochondria and producing heat. It is a non-exercise means of heat production that is often seen during hibernation in animals.
Brown fat cells are most abundant in babies. The reason is to maintain body temperature. This is to protect lives. Adults shiver when exposed to severe cold. This is how babies move their muscles to generate heat, but babies have few muscles and cannot regulate their own body temperature. Babies have a lot of brown fat cells to support life, but they are no longer needed after that and gradually decrease. BMI increases with age. Conversely, brown fat cells decrease, leading to obesity. When brown fat cells, which can be called "fat-burning fat," disappear, it becomes easier to gain weight. It is expected that increasing the number and function of brown adipose tissue will lead to new treatments for type 2 diabetes and obesity.
[UCP-1]
Uncoupling protein is often abbreviated as UCP, which stands for Uncoupling protein. Uncoupling protein (UCP) is a protein in the inner membrane of mitochondria that can dissipate the proton gradient across the membrane before generating energy for oxidative phosphorylation. Five types of UCP-1 to UCP-5 are known in mammals. Instead of producing ATP, the energy is used to generate heat, so the uncoupling proteins perform normal physiological functions such as non-exercise thermogenesis during hibernation. UCP-1 exists only in brown adipocytes, UCP-2 is found in white adipocytes, immune system cells, nerve cells, etc., and UCP-3 is mainly present in large amounts in muscle tissue such as skeletal muscle and heart. Since UCP-3 protein synthesis is markedly reduced in the skeletal muscles of diabetic patients, it is thought to be related to thermogenesis or fat metabolism. When noradrenaline binds to β3 receptors on brown fat cells, UCP-1 is produced, causing uncoupling in the mitochondria and producing heat. Yellow people, including Japanese, often have a genetic mutation in the β3 receptor gene, and while they produce less heat, they are also less able to conserve and consume calories, so they conserve this mutated gene. Sometimes called genes.
Mitochondrial uncoupling protein (UCP) has the function of uncoupling the oxidative phosphorylation reaction in the mitochondrial inner membrane and dissipating energy as heat. Regarding UCP (UCP-1), which is the most representative type of brown adipose tissue, (1) the function of UCP-1 is decreased in obese animals, (2) UCP-1 is increased in animals that do not become obese even if they eat a lot of food. are doing. It is a known fact that mice in which UCP-1 expression is artificially reduced become obese, while mice with high UCP-1 expression become thin. Therefore, if UCP-1 is activated, an anti-satisfaction effect can be expected, and drugs and foods for this purpose are being searched for, and a typical example is an adrenergic receptor agonist specific to adipocytes. In fact, beta agonists promote lipolysis in white adipocytes and at the same time activate UCP-1, converting free fatty acids into heat, ultimately reducing body fat. Unlike mice and the like, adults have only a small amount of brown adipose tissue. However, continued administration of beta-agonists causes normal adipocytes to turn brown and UCP-1 to increase. Furthermore, since UCP-2 and UCP-3, which are homologous to UCP-1, are widely present in human skeletal muscle and white adipose tissue, UCP, including these proteins, is considered to be one of the anti-obesity target molecules. There is.
[PGC-1α]
Involvement of PGC-1α in the promotion of glucose metabolism by exercise: In skeletal muscles that have undergone a certain amount of continuous exercise, the number of intracellular organelles called mitochondria increases and fat burning becomes active, leading to an increase in the amount of fat in the blood. Sugar metabolism becomes active due to an increase in the sugar transporter GLUT4, which takes glucose (blood sugar) into skeletal muscles. PGC-1α, a substance that controls gene transcription, has the function of promoting mitochondrial synthesis and increases GLUT4 in experiments using cultured skeletal muscle cells. PGC-1α is also present in skeletal muscle, and its amount increases with exercise, so it is thought that an increase in the amount of PGC-1α caused by exercise is linked to changes in the properties of skeletal muscle. PGC-1α is a coactivator of PPAR-α, PPAR-γ, and other transcriptional regulators. It is expressed in muscle, liver, and brown adipose tissue. In the liver, expression increases during fasting to promote gluconeogenesis, and in brown adipose tissue, it regulates transcriptional programs related to thermogenic adaptation. Introducing PGC-1α into white adipocytes causes brown adipocyte-like changes such as enhanced mitochondrial biogenesis and increased expression of UCP-1. Full-length PGC-1α is 113 kDa and is induced in brown adipose tissue by cold exposure, in the liver and kidney by fasting, and in skeletal muscle by exercise.

[PPAR-α]
PPAR-α is a member of the nuclear receptor superfamily. To date, three subtypes have been reported: α, γ, and δ (β). The name comes from the fact that the first α subtype (PPAR-α) discovered was activated by fibrate drugs, which are peroxisome proliferators. It is said to be a group of transcription factors that are closely involved in the intracellular metabolism of hydrocarbons, lipids, proteins, etc. and cell differentiation. Both subtypes form heterodimers with the retinoid X receptor (RXR) and bind to the PPAR response element (PPRE).
PPAR-α is strongly expressed in the liver, brown adipose tissue, heart, and kidney, and is activated using physiological ligands such as free fatty acids, leading to a decrease in blood triglyceride concentration. Exogenous ligands include so-called fibrate drugs such as bezafibrate and clofibrate. Most of the target genes are lipid metabolism-related genes, which are the main targets of hypertriglyceridemia-improving drugs.
[Prdm16]
Prdm16 is a transcription factor that plays an important role as a switch for inducing differentiation into brown adipocytes and beige adipocytes. Prdm16 forms a complex with C/EBPβ and has transmethylation activity, thereby inducing differentiation of Myf5-positive cells into brown adipocytes. Lysine methyltransferase EHMT1 was identified as the only histone methyltransferase responsible for the transmethylation activity of the Prdm16 transcription complex. The Prdm16/EHMT complex plays a role in suppressing skeletal muscle-related gene expression in mouse BAT and activating a genetic program for differentiation of precursor brown adipocytes into brown adipocytes. Furthermore, Prdm16 suppresses the expression of white fat-related genes such as resitin and induces beige fat-related gene programs, and plays a very important role as a differentiation switch from preadipocytes to beige adipocytes. ing.
[Tfam]
Mitochondrial transcription factor A
scription factor A: TFAM) was purified and cloned by Clayton et al. as a transcription factor for mitochondrial DNA. TFAM is a protein belonging to the high mobility group (HMG) family of proteins, and like many HMG family proteins, can bind to DNA non-specifically to DNA sequences. There is a correlation between the amount of change in TFAM and the amount of mitochondrial DNA, and the replication of mitochondrial DNA is dependent on transcription, so the expression amount of TFAM is used as a substitute for evaluating mitochondrial function. Mitochondria are responsible for most ATP production through oxidative phosphorylation and are the center of energy metabolism in living organisms.
[Adipocytokine]
A general term for physiologically active substances secreted from fat cells.
[Leptin]
Hormone secreted from fat cells. It has the function of suppressing appetite and activating energy metabolism.
[HbA1c]
Glucose is bound to hemoglobin (Hb) present in red blood cells. The lifespan of red blood cells is approximately four months, during which time they circulate around the body and glucose binds to hemoglobin. The higher the amount of glucose in the blood, the higher the HbA1c value (hemoglobin A1C), and the HbA1c value reflects the state of blood sugar control over the past 1 to 2 months.
[GA]
Glycoalbumin is a glycated protein that reflects blood sugar status. It is used as an indicator of blood sugar control over the past 1 to 2 weeks.
[Adiponectin]
It is a secretory protein secreted from fat cells. The concentration in the blood is an order of magnitude higher than that of general hormones, reaching the μg/ml order. Its actions include promoting sugar uptake that is not mediated by insulin receptors, burning fatty acids, increasing insulin receptor sensitivity by reducing intracellular fatty acids, and increasing insulin sensitivity by activating AMP kinase in the liver. It has a variety of effects, including inhibiting arteriosclerosis, anti-inflammation, and inhibiting myocardial hypertrophy. It is a good adipocytokine.
[TNF-α]
Adipose tissue secretes inflammatory cytokines, and TNF-α inhibits glucose uptake into cells and reduces sensitivity to insulin. It has also been reported that TNF-α promotes the production of fatty acids in adipocytes and hepatocytes and causes antiglycerinemia mainly through TNFR1. One of the adipocytokines (physiologically active substances) secreted by fat cells, which has the effect of suppressing the action of sugar in muscles, adipose tissue, and the liver. It increases when you are obese, increasing your risk of diabetes and arteriosclerosis. It is secreted from fat cells and is well known as a cytokine that induces insulin resistance.
[MCP-1]
It is secreted from the inflammatory layer (vascular endothelial cells, adipocytes), induces the migration of monocytes, differentiation into macrophages, and expression of oxidized LDL receptors, and is an important factor in the formation of arteriosclerosis. By administering D-psicose for 3 months, the MCP-1 concentration was significantly reduced, suggesting that D-psicose has an anti-arteriosclerotic effect.
[Oxidized LDL receptor]
The LDL (low-density lipoprotein) receptor family is a multifunctional protein that controls the intracellular uptake of various ligands, including LDL, and signal transduction. When LDL is oxidized by oxidizing substances such as free radicals and becomes oxidized LDL, it is not recognized by normal LDL receptors, but is recognized by scavenger receptors of macrophages, and is taken up indefinitely, leading to foam formation by macrophages. became clear.

(Example 1)
We investigated the effects of administration of D-allulose, a type of rare sugar, on brown adipose tissue (BAT). In the experiment, 6-week-old mice were used, divided into 3 groups with 5 mice each, and treated as described below for 8 weeks.
1. Protocol:
(1) Group that eats normal food and drinks water (Normal Food)
(2) Group eating high-fat food and drinking water (HFD)
(3) Group receiving a high-fat diet and drinking water containing 2% D-allulose (HFD+2% D-allulose)

2. Evaluation item:
(1) Changes in body weight,
Changes in blood glucose levels (2) Size and weight of subscapular brown fat cells (3) Pathological evaluation of morphological changes in brown fat cells (4) UCP for brown fat cell activation -1, Evaluation of expression levels of PPAR-α and PGC-1α

3. result:
1) Figure 1 shows the results of changes in body weight and blood sugar levels.
Weight increased in the group fed a high-fat diet. A decrease in body weight was observed in the high-fat diet and D-allulose groups (HFD+2% D-allulose) compared to the high-fat diet group. Regarding the changes in blood sugar levels, the high-fat diet group showed high blood sugar levels, but the high-fat diet and D-allulose groups showed a decrease in blood sugar levels.
2) The size and weight of subscapular brown fat cells, and the results of body weight are shown in Figure 2.
Brown fat cells decreased in the group fed a high-fat diet. An increase in brown adipocytes was observed in the high-fat diet and D-allulose groups compared to the high-fat diet group.

3) The results of pathological evaluation of morphological changes in brown fat cells are shown in Figure 3.
Histological staining also showed fattening in the high-fat diet group, but improved to a normal state in the high-fat diet and D-allulose groups (HFD + 2% D-allulose).
4) The results of evaluating the expression levels of UCP-1, PPAR-α, and PGC-1α for brown adipocyte activation are shown in FIGS. 4, 5, and 6, respectively.
In the high-fat diet and D-allulose groups (HFD + 2% D-allulose), enhanced brown adipocyte marker gene expression was observed compared to the high-fat diet group.

summary:
As shown in Figure 1, regarding the effect of D-allulose on mouse body weight, high-fat diet (HFD) increased body weight over 8 weeks of observation. When the HFD group was given water containing 2% D-allulose, a decrease in body weight was observed. Blood sugar levels were also measured over time during the 8 weeks. In the HFD group, blood sugar levels were higher than in the normal group. Blood glucose levels in the HFD+D-allulose group were lower than in the HFD group.
Figure 2 shows the size, weight, and body weight of subscapular brown fat cells in the experiment of this example using 6-week-old mice, and a diagram showing the results of pathological examination of brown fat tissue in three groups. As shown in No. 3, fat deposits were observed in BAT in the HFD group compared to the normal diet group, which turned into white fat. On the other hand, in the HFD group + 2% D-allulose group, the brown adipose tissue was morphologically comparable to that in the normal diet group, and it was found that the brown adipose tissue that had turned into white fat had recovered to normal brown adipose tissue. .
As shown in Figure 4, which shows the activation of brown adipocytes by the expression level of UCP-1, UCP-1 was increased in the HFD group. It was predicted that ingesting a large amount of fat would induce UCP-1 as a body defense response, leading to calorie consumption. Administration of D-allulose further induced UCP-1. When considered in conjunction with pathological tissue, the induction of UCP-1 in the HFD group is a response to a pathological situation, and in the HFD + 2% D-allulose group, brown adipose tissue is induced due to the normal morphology. It is presumed that this is the activation of UCP-1 as a result of differentiation and proliferation.
As shown in Figure 5, which shows the expression level of PPAR-α indicating the activation of brown adipocytes, PPAR-α is an indicator of the activation of brown adipocytes, and PPAR-α was significantly lower in the HFD group than in the normal group. was induced, but this seems to be a biological reaction due to excessive lipid intake. In the HFD group + 2% D-allulose group, PPAR-α was more strongly induced by the intake of D-allulose, suggesting that brown fat cells were activated.
As shown in Figure 6, which shows the activation of brown adipocytes by the expression level of PGC-1α, PGC-1α was induced in the HFD group compared to the normal group, but this was probably due to excessive lipid intake. It seems to be a biological reaction. In the HFD + 2% D-allulose group, PGC-1α was more strongly induced by D-allulose intake, and the induction of PGC-1α, which is a coactivator of PPAR-α, is due to UCP-1, which is located in the downstream region. This is thought to be one of the reasons why brown fat cells are activated.
It is thought that administration of D-allulose stimulated the activation of brown fat cells and contributed to weight loss by enhancing thermogenesis, fat burning, and metabolism. As for future prospects, it is possible that by ingesting D-allulose, it is possible to acquire a constitution that is ``easy to lose weight.''

(Example 2)
We investigated the effects of the rare sugar D-psicose (D-allulose) on type 2 diabetic patients.
[Method]
<Selection criteria>
Type 2 diabetic patients who do not receive sufficient effects from any of the following treatments:
(HbA1c: 6.5% or more) (borderline diabetes)
1) Diet/exercise therapy only 2) Drug therapy in addition to diet/exercise therapy <Exclusion criteria>
1) Patients with contraindications to D-allulose (D-psicose) administration 2) Patients participating in other clinical trials 3) Women who are pregnant, giving birth, breastfeeding, or who may become pregnant 4) HbA1c is 8% or higher 5) Patients with severe renal dysfunction (serum creatinine level 1.5 mg/dl or more) 6) Patients with other serious complications <Experiment implementation method>
D-allulose (D-psicose) powder: 1 stick 5g/packet (manufactured by Rare Sweet Co., Ltd.)
The drug was orally administered at a dose of 5 g three times a day.
General findings and blood tests were compared before and 12 weeks after administration in 12 type 2 diabetic patients (4 males, 8 females) who could be observed for 3 months. (Wilcoxon signed rank sum test)
The implementation complies with the Declaration of Helsinki and the ethical guidelines for clinical research and is approved by the Ethics Committee of Kagawa University School of Medicine.

<Consideration>
In this study, by administering D-allulose (D-psicose) for 3 months, a significant decrease in body weight was observed after 3 months.
Regarding various adipose tissue-derived physiologically active substances (adipocytokines) and their effects, no significant changes were observed in leptin and adiponectin during the 3 months of D-psicose administration.
As is well known, TNF-α is secreted from adipocytes and is famous as a cytokine that induces insulin resistance. It is also known to suppress adiponectin, a good adipocytokine. It is presumed that the administration of D-psicose in this example lowers TNF-α, which improves insulin resistance and improves blood sugar control.
Furthermore, MCP-1 is secreted from the inflammatory layer (vascular endothelial cells, adipocytes), induces monocyte migration, macrophage differentiation, and oxidized LDL receptor expression, and is an important cytokine that forms arteriosclerosis. . Administration of D-allulose (D-psicose) for 3 months significantly reduced MCP-1 concentration, suggesting that D-allulose (D-psicose) has anti-arteriosclerotic effects. .

<summary>
(1) D-allulose (D-psicose) was orally administered at a dose of 5 g three times a day to 12 type 2 diabetic patients (4 males, 8 females), and general findings were observed 12 weeks before and after administration. , blood tests were compared.
(2) No significant differences were observed in HbA1c, GA, leptin, and adiponectin.
(3) TNF-α and MCP-1 were significantly decreased before and after administration. In particular, TNF-α decreased significantly two months after administration.
(4) Body weight decreased by an average of 1 kg three months after administration.
(5) D-allulose (D-psicose) may be useful for administration to type 2 diabetic patients.

(Example 3)
Similar to Example 1, regarding the effect of D-allulose administration on brown adipose tissue (BAT), it was found that induction of differentiation of beige adipocytes led to weight loss and body fat reduction. We investigated the effects of
(Purpose) To confirm the effect of administration of D-allulos on brown adipose tissue in mice. We will also investigate the differentiation of beige adipocytes using changes in the expression of UCP-1 and other proteins as markers.
1. Protocol:
Each group consists of 5 mice and is divided into 2 groups. Both groups of 8 week old mice are fed a high fat diet (HFD). Four weeks after HFD loading, group 1 continued on HFD. In another group, in addition to Hfd, D-allulos (0.2 mg/g body weight/day) was administered into the stomach using a probe. Body weight, food intake, water consumption, and blood sugar levels will be monitored until the 10th week.
2. Results 1) Figure 10 shows the results regarding body weight increase during high-fat diet (HFD) loading.
Weight gain in five mice each was observed when fed a normal diet and a high-fat diet (HFD). When loaded with a high-fat diet (HFD), a clear weight gain is observed compared to when fed a normal diet.
2) Figure 11 shows the change in body weight when D-allulose was administered to mice fed a high-fat diet (HFD).
After 4 weeks of high-fat diet (HFD) loading before administration of D-allulose, no difference in weight gain was observed between the two groups.
A significant difference in body weight between the two groups was observed from 2 weeks after the start of D-allulose administration (6 weeks after starting the high-fat diet).
In the D-allulose administration group, the increase in body weight was significantly reduced from the second week after the start of administration.
3) Regarding the transition of fasting blood sugar level, the results are shown in FIG. 12.
It shows changes in body weight when D-allulos was administered to mice loaded with a high-fat diet (HFD). A decrease in fasting blood sugar level was observed from 3 weeks after the start of D-allulos administration (6 weeks after starting the high-fat diet).
4) The results of the blood glucose tolerance test after 10 weeks are shown in FIG. 13.
Glucose was administered intraperitoneally, and blood sugar levels were measured 0, 15, 30, 60, 90, and 120 minutes later. In the D-allulose administration group, blood glucose levels after 90 and 120 minutes were significantly lower than in the control group.
5) As shown in Figure 14, there were no significant changes in food intake (right panel) or water intake (left panel) during the study period.
6) The results of the study regarding the weight and area of brown adipocytes (BAT) when D-allulos was administered to mice loaded with a high-fat diet (HFD) are shown in FIG. 15.
In the D-allulose administration group, an increase in the weight and area of brown fat cells (BAT) was observed, and the same results as in Example 1 were obtained.
In the morphological study of brown adipose tissue (BAT), similar to Figure 3 of Example 1, the histology of brown adipose tissue (BAT) when consuming a normal diet was significantly different from that when consuming a high-fat diet (HFD). The histological image of the brown adipocytes (BAT) of the challenged mice showed that fat was deposited on the whole, and fatification was progressing.On the other hand, when the mice were loaded with a high-fat diet (HFD) and were given 2% D-allulos in their drinking water. The histological image of brown fat cells (BAT) in the group was almost the same as that in the normal diet group, and no fat deposition was observed in the high-fat diet (HFD) group.

(About adipose tissue)
Here, the current way of thinking about adipose tissue (non-patent document 3) will be explained with reference to FIG.
Beige adipocytes, or beige fat cells, were isolated in 2012 as a third type of fat cell by Dr. Bruce Spiegelman's research team at Harvard Medical School's Dana-Farber Cancer Institute.
Fat cells include white fat cells, which store fat, and brown fat cells, which burn fat and produce heat. Brown fat cells express a large amount of a protein called uncoupling protein 1 (UCP-1), which generates heat, burns fat, and converts it into energy. Beige adipocytes, like white adipocytes, express very low UCP-1, but UCP-1 is highly expressed by stimuli such as cold. At that time, these adipocytes begin to produce heat like brown adipocytes, and white adipocytes take on a brown-like appearance. Brown adipocytes and beige adipocytes are specialized adipocytes that generate heat in response to cold exposure, contributing to maintaining body temperature in cold environments. The thermogenic and energy consuming activities of these fat cells are expected to be useful not only for regulating body temperature but also for preventing obesity and metabolic diseases. Brown adipocytes and beige adipocytes express UCP-1 and have thermogenic ability in common, but it has been found that their cell origins and functional control mechanisms are different. Classical brown adipocytes are developed in small rodents, especially hibernating animals, and exist as brown adipocyte clusters between the shoulder blades, in the axilla, and around the kidneys. While brown adipocyte differentiation and tissue formation are completed during the fetal period, beige adipocyte differentiation is induced in response to stimuli such as exposure to a cold environment, and disappears when the stimulus is removed. This inducibility and plasticity can be said to be the most important feature compared to the "existing type" of brown adipocytes and white adipocytes, which continue to exist from the time of development.
Looking at their functional characteristics, unlike white adipocytes, which store surplus energy as neutral fat, brown adipocytes and beige adipocytes generate heat by uncoupling oxidative phosphorylation by UCP-1. . In these respects, beige adipocytes can be said to be similar to brown adipocytes.
In recent years, research using positron imaging (FDG-PET) has revealed that a certain amount of brown adipose tissue (BAT) exists in humans as well. The locations of BAT in humans include the interscapular region, the perirenal region, the supraclavicular region, the lower axilla, and the paravertebral region. Recent studies have suggested that BAT in adults is mainly composed of beige adipocytes. This is also supported by the fact that adult BAT activity is minimal in summer and maximal in winter, ie, inducible and plastic. A negative correlation is observed between the amount of change in BAT activity and the amount of change in body fat. This result revealed that human BAT (beige adipocytes) is an effective stimulation target for reducing obesity.

7) The results of examining the expression of UCP-1 mRNA in beige adipocytes and brown adipocytes in mice fed a high-fat diet (HFD) and administered D-allulose are shown in FIG. 17.
The drawing on the left shows that the expression of UCP-1 is significantly increased in beige adipocytes by administration of D-allulose. This indicates that beige adipocytes are induced to differentiate by D-allulose.
The drawing on the right shows that UCP-1 expression is increased even in classical brown adipose tissue.
Administration of D-allulos may contribute to weight loss through induction of beige adipocytes and activation of brown adipocytes.
8) FIG. 18 shows the results of examining the expression of UCP-1 protein in beige adipocytes and brown adipocytes in mice fed a high-fat diet (HFD) and administered D-allulose.
The drawing on the left shows that in beige adipocytes, the expression of UCP-1 protein is significantly increased by administration of D-allulose. This indicates that beige adipocytes are induced to differentiate by D-allulose.
The drawing on the right shows that UCP-1 protein expression is increased even in classical brown adipose tissue.
Administration of D-allulos may contribute to weight loss through induction of beige adipocytes and activation of brown adipocytes.
9) Details of induction of beige adipocytes and activation of brown adipocytes by D-allulos administration include genes involved in heat production, UCP-1 and Prdm16, and genes reflecting the function of mitochondria, an organelle involved in heat production. , Pgcl-α, Tfam, and the expression of PPARγ, a gene related to adipose differentiation, were investigated, and the results are shown in FIG. 19.
The drawing on the left shows that in the examination of beige adipocytes, the expression of UCP-1, Prdm16, Pgcl-α, Tfam, and PPARγ was increased, indicating that beige adipocytes were strongly induced.
The drawing on the right shows that the expression of UCP-1, Prdm16, and Tfam was increased in brown adipocytes. It is presumed that these are involved in the activation of brown fat cells.

10) Figure 20 shows an experiment using cells (establishing a cell line derived from a single clone from adult supraclavicular BAT and using a model for the study of differentiation induction into beige adipocytes). -Protocol and WAT-Protocol are drawings explaining protocols for inducing beige adipocytes.
FIG. 21 is a diagram illustrating a protocol in which D-allulose is added to the BAT-protocol and WAT-protocol, which induce beige adipocytes in experiments using cells.
FIG. 22 shows by fat staining that addition of D-allulose to the differentiation induction protocol promotes induction into beige adipocytes.
Addition of D-allulose (right) promotes оil-red-O (fat staining).
FIG. 23 examines the expression of various markers as an evaluation of beige adipocytes induced by the differentiation induction protocol. The WAT-protocol has a stronger effect on the induction of browning marker genes (UCP-1, Pgc-1α, cox8b). Using the WAT-protocol, D-allulose enhances the expression of Prdm16, Pgc-1α, and PPARγ.
In both differentiation protocols, D-allulose enhanced PPARγ expression, suggesting that D-allulose promotes adipogenesis.
That is, in the D-allulose administration group, all markers significantly increased, and the process of beige adipocyte induction was significantly promoted.
In FIG. 24, the expression of various markers is examined as an evaluation of beige adipocytes induced by the protocol. We are investigating UCP-1 and PPARγ in the presence and absence of D-allulose. It was shown that the presence of D-allulose increased UCP-1 and PPARγ, promoting the induction of beige adipocytes.
Compared to the BAT-protocol, the WAT-protocol has a stronger effect on the induction of UCP-1 protein expression. Using the WAT-protocol, D-allulose slightly increases UCP-1 protein expression.
3. Conclusion: Weight loss is observed with administration of D-allulose. This weight-reducing effect is thought to be due to induction of differentiation of beige adipocytes and activation of brown adipocytes.

By ingesting the brown fat cell activator of the present application containing D-allulose, it activates brown fat cells and beige fat cells and promotes energy consumption throughout the body, thereby reducing fat mass and eventually preventing obesity. It is hoped that this will be resolved. In fact, in humans, intake of D-allulose (15g per day) significantly reduced body weight after 3 months.In humans, D-allulose also contributes to the proliferation of brown adipose tissue and the acquisition of a ``lean body.'' In addition, as we age, metabolism generally decreases and the body feels colder, but D-allulose intake can prevent and improve the body's coldness by increasing energy metabolism (thermogenesis). It is also expected that D-allulose will induce differentiation of brown-like adipocytes (beige adipocytes) in white adipose tissue due to its brown adipocyte activation effect.

Claims (12)

An agent for morphologically increasing the weight and area of brown adipocytes or beige adipocytes whose functions are activated by D-allulose, which contains D-allulose as an active ingredient. The brown adipocytes or beige adipocytes according to claim 1, wherein the functionally activated brown adipocytes or beige adipocytes are brown adipocytes or beige adipocytes in which the expression level of an activation indicator is increased. A morphological increaser in weight and area. The brown adipocytes or beige adipocytes in which the expression level of the activation indicator is increased are brown adipocytes or beige adipocytes in which UCP-1 expression is promoted and the expression level is increased. An agent for morphologically increasing the weight and area of brown fat cells or beige fat cells. The brown adipocytes or beige adipocytes in which the expression level of the activation indicator is increased are brown adipocytes or beige adipocytes in which UCP-1 expression is enhanced and the expression level is increased. An agent for morphologically increasing the weight and area of brown fat cells or beige fat cells. An energy consumption promoter containing D-allulose as an active ingredient, which is an agent for morphologically increasing the weight and area of brown fat cells or beige fat cells whose expression levels of activation indicators are increased by D-allulose. The brown adipocytes or beige adipocytes in which the expression level of the activation indicator is increased are brown adipocytes or beige adipocytes in which energy consumption is promoted due to the expression of genes involved in thermogenesis, and the expression level is increased. Item 5. The energy consumption promoter according to item 5. A UCP-1 expression enhancer containing D-allulose as an active ingredient, which is an agent for morphologically increasing the weight and area of brown adipocytes or beige adipocytes in which the expression levels of activation indicators are increased by D-allulose. A composition for morphologically increasing the weight and area of brown adipocytes or beige adipocytes, comprising the agent according to any one of claims 1 to 7. A composition for morphologically increasing the weight and area of brown adipocytes or beige adipocytes in which the expression level of an activation index is increased by D-allulose, which contains D-allulose as an active ingredient. The brown adipocytes or beige adipocytes in which the expression level of the activation indicator is increased are brown adipocytes or beige adipocytes in which energy consumption is promoted due to the expression of genes involved in thermogenesis, and the expression level is increased. Item 9. The composition for morphologically increasing the weight and area of brown adipocytes or beige adipocytes according to item 9. According to claim 9, the brown adipocytes or beige adipocytes in which the expression level of the activation indicator is increased are brown adipocytes or beige adipocytes in which the expression of UCP-1 is enhanced and the expression level is increased. A composition for morphologically increasing the weight and area of brown adipocytes or beige adipocytes. The food composition for morphologically increasing the weight and area of brown adipocytes or beige adipocytes according to any one of claims 9 to 11. Composition for use.